Development of Methods and Techniques to Visualize Mechanical Tension in Embryos Using Genetically Encoded Fluorescent Mechanosensors

Abstract

Lately, the growing body of quantitative data has provided evidence of the importance of mechanical forces in embryogenesis. The study of spatial and temporal distribution of mechanical tension in the course of embryogenesis is one of the most important problems of modern developmental biology. Development of genetically encoded fluorescent mechanosensors allowed their application in an intravital study of mechanical tension in developing embryos via noninvasive techniques. The possibility of applying fluorescent mechanosensors based on vinculin and C-cadherin to visualize mechanical tension in tissues of Gallus and Xenopus embryos was studied. The methods to express and detect these proteins, as well as process the resulting images, were elaborated. The best results were obtained using Xenopus embryos and the vinculin-based mechanosensor.

Keywords:

Notes

ACKNOWLEDGMENTS

The work was supported by the Russian Foundation for Basic Research (project no. 15-04-06310). Works on cloning of plasmid constructs were supported by the Russian Science Foundation (project no. 14-50-00131).